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In Vitro Regeneration of Chickpea (cicer Arietinum L.)

M. M. Khatun
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

M. R. Kabir
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

D. Khanam
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

M. A. Y. Akhond
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

Abstract/ Executive Summary:

Mature embryo and cotyledon pieces of BARl Chola 7 were used for in vitro regeneration. Four different concentrations of 2, 4-D (T₁, T₂, T₃, & T₄) were used for callus formation and shoot development. The highest percent of explant (100%) produced callus in T₁ & T₄ treatments from isolated mature embryo explant. In case of cotyledon explant, 100% callus formation was observed in all the treatments. In shoot regeneration T₂ performed better.

Keywords: Chickpea, In vitro regeneration, Shoot regeneration

Location: Pulse Research Centre (PRC) Joydebpur, Gazipur, Bangladesh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Chickpea

Objectives:

Objective of this study was to develop an efficient regeneration protocol for Barichola which will be used for future for the development of Chickpea transgenic with desired traits.

Methodology:

Chickpea seeds were collected from Pulse Research Centre (PRC) Joydebpur, Gazipur. Healthy seeds were surface sterilized with detergent for 10 min and washed with running tap water for 15 min. Then the seeds were sterilized with 0.1% HgCl₂ with 1-2 drops of Tween 20 for 10 min. and washed three to four times with sterile distilled water. After that the seeds were soaked overnight in sterile distilled water. For callus formation, different concentrations of 2, 4-D were used in MS medium. For shoot initiation, explants were cultured on MS medium supplemented with various hormonal concentrations namely BAP, Kn, NAA, TDZ and GA₃. All media had 3% sucrose and 0.8% agar. The pH of all media was adjusted to 5.8 before autoclaving. The media were autoclaved at 121⁰C and 1.06 kg cm² pressure for 20 minutes. The seeds were split open and mature embryo were separated and put on callus induction medium. Single Cotyledon pieces were also placed on callus induction medium. For regeneration of shoots, internode and leaf explants from in vitro grown seedling were also used. All cultures were incubated at 25 ± 1⁰C and kept in 16/8 hours light/dark photoperiod with a light intensity of 2000 lux under cool white fluorescent tubes.

Source of Information: Annual Research Report (2013-2014), Biotechnology Division, BARI, Gazipur, Bangladesh

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In embryo explant, the highest percent (100%) explant produced callus in T₁ & T₄ treatments. In the above two treatments, callus quality was also found very well. On the other hand, 93% explant produced callus in T₂& T₃ treatments. In cotyledon explant, 100% callus formation was observed in all treatments. Callus quality was also very good in T₁, T₃, & T₄ treatments. Calli were then transferred to four regeneration media (T₁, T₂, T₃, & T₄) where calli were turned to greenish in colour and shoot started to initiate. Among the four treatments, T₂performed better.

Knowledge Management: Report/Proceedings